Estrogenic substances



Patented Nov. 1, 1949 ESTROGENIC SUBSTANCES Ralph C. Tallman, Morristown, N. J., and Alfred H. Stuart, New York, N. Y., assignors to Schieffelin & 00., New York, N. Y., a corporation of New York No Drawing. Original application May 30, 1944, Serial No. 538,089.. Divided and thisapplication November 10, 1945, Serial No. 627,973

4 Claims.

This application is a division of co-pending application Serial Number 538,089,.filed May 30; 1944, now Patent No. 2,400,033 dated Mayv '7, 1946, which in turn is a continuation-in-part-of application Serial Number 363,669, filed October 31, 1940, now abandoned.

This invention relates to estrogenic substances, i. e., substances possessing properties similar to those possessed by the female ovarian follicular hormone.

Estrogenic substances are those which bring about the physiological changes known asestrusor heat periods in female animals, or the follicular phase of. the menstrual cycle in human females.- Ordinarily-such substances are secreted by the ovaries and the physiological. changes brought about by the normal functioning of these ovarian secretions in conjunction with to substitute for the absent or diminished natural secretions.

For industrial elaboration, estrogenic' substances may be obtained from natural sources, such as the excretions of pregnant female mammals. For example, the urine of pregnant female horses may be collected and subjected to various processes designed to concentrate and purify the estrogenic substances contained therein, and thus to isolate them in a condition suitable for therapeutic administration in the manners and for the purposes named above. Such procedures are devious, time-consuming; and expensive.

This invention describes artificial estrogenic substances which may be prepared from simple chemical compounds. When the products described in this invention are employed, no recourse to estrogenic substances from natural discovery that compounds having the following 2 This invention is based on the discovery that estrogenic and bactericidal properties are possessed by compounds having the structure X0 i i OX i-K3 n H H in which each X is H, alkyl, acyl of aliphatic or aromatic type, or an inorganic: ester radical such as SOsI-I or PO3H2 which may be obtained by condensing one of these compounds-having OH terminal groups with an inorganic acid; and R is H or alkyl, at least one R being alkyl; in which the OX groups may be symmetrical or not, and may. bethe same or different; andi'n which each alkyl B may. bathe same or different.

The invention is further based on the further type formulae'are-especially and uniquely beneficial, and.- itis a furtherobjector? the invention to utilize said compounds as estrogenicand bactericidal compounds.

TYPEI in which X is H; alkyl, or acyl, andR is alkyl.

TYPE II H R H ox -Q i l. l

in which X is H, alkyl, or acyl; and R is alkyl.

' TYPE III H R a ox G 0 if 1'1 t in which X is H, alkyl,. or acyl, and each R is alkyl.

TYPE D g-gear in which X is H, alkyl, or acyl, and each R is alkyl.

The OX radical may be in the ortho, meta, or para position in each of the foregoing types, and need not be symmetrically positioned.

Examples of the production of compounds of the above types are as follows:

EXAMPLE 1.PRODUCTION OF TYPE I Specific members of the series illustrated by Type I may be prepared in the folowing manner: 48 grams of p-methoxyacetophenone and 45 grams of anisaldehyde are disolved in 150 cc. of alcohol. The solution is stirred vigorously, and 150 cc. of aqueous sodium hydroxide solution is gradually added in small portions. The product separates slowly from the solution and becomes crystalline on the addition of seed. Stirring is conshown by separation of crystals or by failure of the residue to boil at a constant temperature, the same process may be repeated until reduction is complete. The final product of this step, l,3-di(p-methoxyphenyl) -pentane may be purified by distillation, and is a colorless liquid which boils at 160-165" C. under 1 mm. pressure. It may be converted into the corresponding phenolic product as follows:

2.0 grams of the 1,3-di(p-methoxyphenyl)- pentane are hydrolyzed by dissolving in 6 cc. glacial acetic acid, to which solution 15 cc. of 5'7 aqueous hydriodic acid is added. The reaction mixture is maintained under refiux at the boiling point for 15 minutes, then 4 cc. of glacial acetic acid and 5 cc. of the hydriodic acid solution are added, and refluxing continued for a total of four hours. The mixture is poured into water, the solution made alkaline with excess potassium hydroxide, and filtered with the aid of charcoal. The phenolic product thus obtained (see Step IV below) is precipitated by acidification of the filtrate, extracted with ether, and, after the evaporation of the ether, distilled slowly at 150 C. under high vacuum. The distillate is a colorless glass-like product which solidifies on long standing. This compound is 1,3-di-(p-hydroxyphenyl) -pentane, where R is the ethyl radical.

Pnontlcrron or TYPE I Step I Step II cmo-O g-ormcn-O-ocn. EtMgBr fiukQe-omouOoom Step III H; oruoOcm-om-on 00H,

(reduction) {1 Step IV omo-Qom-om-oH-Ooom HOG-CHr-CIIr-CH-OOH it (hydrolysis) 1] nesium bromide in 450 cc. dry ether is maintained at 10 C. and vigorously stirred while 20 grams of the chalcone is added in small quantities. The cooling bath is then removed, and stirring continued for two and one-half hours at room temperature. The reaction mixture is poured into 1200 cc. ice water containing 150 cc. hydrochloric acid, after Which'the ether layer is-separated and the aqueous layer extracted once with ether. The ether is evaporated from the combined solutions and the product which remains is purified by distillation under high vacuum and crystallization from alcohol. The white crystals of the product, 1,3-di(p-methoxyphenyl) -pentanone-1 (Where R is the ethyl group) have a melting point of C. i

This compound may be converted into 1,3-di(pmethoxyphenyl) -pentane as follows:

9 grams of the pentanone are mixed with 60 grams of amalgamated zinc, 45 cc. water, cc. hydrochloric acid, 35 cc. of toluene, and 2 cc. acetic acid, and the reaction mixture heated under reflux at the boiling point for twentyeight hours (see Step III below). It is then cooled and extracted with ether, the ether layer separated, and all solvents removed therefrom by 7 of hydrogen chloride has ceased, after which it is evaporation. If reduction is not complete, .as

.Any product corresponding to Type I can be produced by following the basic outline of the process and equations set forth herein above, using an alkyl magnesium halide having the desired number of carbon atoms in its alkyl group. The condensation can also be carried out in acid medium, using a mineral acid as the condensing agent.

EXAMPLE 2.PRODUCTION OF TYPE II Specific members of the series illustrated by Type II may be prepared in the following manner: 16 grams of anisaldehyde and 22 grams of pmethoxybutyrophenone are thoroughly mixed, and a stream of dry hydrogen chloride gas bubbled into the mixture until the total weight is increased by 3.5 grams. The reaction mixture, which rapidly becomes very dark red, is shaken until it becomes pasty, due to the separation of the product, and is then allowed to stand for fifteen hours. The entire mixture is dissolved in benzene and the benzene solution extracted with water, with sodium carbonate solution and again with water. The benzene is then evaporated and the residue maintained at C. until evolution a 2. "and: R11" mayv beasthe; same" or diflerent 'alkyl 'i radicals. In general any desiredcompiound'corresponding'to this type formula-may be made by using, in Step'I, a keton'ehaving the proper alk'yl radical and, in 'Step II a magnesium bromide compound having the proper alkyl radical.

. Since the carbon atoms to which the alkyl radicals-are .:attached are asymmetric in this type of compound, the product produced .may be a single racemate or a mixture of racemates and the individual racemates of such mixture may be separated by any suitable method, a preferred method being disclosed in our copending application" Serial No. 538,090 filed May 30, 1944, now l=.'atent;;l\lo.v 2,400,034, dated May 7, 1946. For exam'ple;when R and R1 are'ethyl groups, the intermediate compound produced as shown in Step II is in the former two'racemicmixtures, one liquid and one solid. I @These mixtures may be separated by dissolving them in a solvent such asalcohol and crystallizing the solid racemie mixture from the solution. "By' evaporating the solvent from the motherliquid, the liquid racemic mixture may be recovered. Either the solid or the liquid racemic mixture may-'the'nbe subjected to the reactions shown in'Steps" III and IV. As also stated in that application; an individual racemic mixture 8 The etherdaye'r is separated. and the ether removed. I The residue is subjected to vacuum distillation, and is obtained as a colorless oil which has a boiling point of 183184 C. at 2 mm. and which solidifies to a solid compound melting after purification at 87 C. During this distillation water is evolved and the resulting product is '3,5-di(p-methoxyphenyl) -heptene-2, where R and R1 are ethyl radicals (see Step in below). This is then treated according to Step IV as follows: 4 grams of the 3,5-di(p-methoxyphenyl) -heptone-2 is dissolved in 70 cc. of glacial acetic acid and 0.04 gram of platinum oxide catalyst added. The container is then attached to a machine for catalytic hydrogenation, the hydrogen applied, and the mixture shaken until one molecular equivalent of hydrogen has been consumed. The mixture is then decanted from the catalyst, diluted with water, neutralized with NaOH, and the product extracted with ether. The ether is evaporated at room temperature, and the residue distilled under reduced pressure. The product thus obtained (see Step IV below) is a colorless oily liquid and is 3,5-di(p-methoxyphenyl)-heptane, where R and R1 are ethyl radicals; and may be converted into the corresponding phenolic compound as in Example 1 (see Step V below).

Paopucrrort or TYPE IV Step1- omQOE eHH onto E-crhCIFOoon,

Step If Step III 1 ontimtc in go-om oi two stereoisomeric terms .ofthis type of com-.- pound may have gre 3 estrogenic potency than Step I below). This is then treated as follows:.,.

Following exactly the same directions as de-" scribed inExample 1, Step II, the p,p'-dimethoxy chalcone is converted into 1,3 -di(pmethoxy- 65 phenyD-pentanone-l (see Step II below. This is then treated according to Step III as followszl il. 1-

A solution of 10 grams of the pentanone in 130 cc. dry ether may be added slowly with vigorous stirring to a's'olution of 8.9 grams of ethyl magnesium bromide in 60 cc. dry ether. After the addi tion of the pentanone has been completed, the

and II below. This may be converted into other mixture is warmed-pn-a steam bath 'for five hours,

after whichthemixture is poured into .a mixture.

: R and R1 may be 'thesame 'or diiferent alkyl radicals. ,.In general,- any compound conforming to this type formulamay be made by using in Steps II and III magnesium bromide compounds having alkyl radicals of the constitution desired in the final product.

Since the carbon atoms to which the alkyl radicals are attached are asymmetric in this type of compound, the product may be a single racemate 013a mixture of racemates and the individual racemates of such mixture may be separated.

EXAMPLE 5.-PRODUCTION OF TYPE V Specific members of the series illustrated by Type V may be prepared in the following manner: The first two steps in this process are identical inallrespects with the first two steps in Example 3 for the production of 1,3-di(p-methoxyphenyl) -2-ethyl pentanone-l (see Steps I and productsas'follows 5 I 8.0 gramsof the ethyl pentanone obtained in o: 700 cc. ice water arid 100 cc. hydrochloric acid. 75 Step II are dissolved in dry ether and added slowsubjected to distillation under reduced pressure; After a small amount of lower boiling forerun, which is discarded, the desiredproduct distills at 190-200? C. under 1 mm. pressure. The product, 4,4 -dimethOXyFa-ethyl chalcone, where R is the ethyl group, (see Step I below) is a bright yellow oil.

grams of the 4,4-dimethoxy-u-ethyl chalcone from the previous procedureis dissolved in cc. of absolute alcohol, 1 gram of a copperchromium oxide catalyst added, and the mixture submitted to high-pressure hydrogenation. at 220 and about 150 atmospheres pressure for 5 hours in an appropriate machine. After cooling, the hydrogen pressure is released.- and the solution removed from the hydrogenation apparatus andfiltered free of the catalyst. Oncooling thefiltrate, the product, 1,3-di--(pemethoxyphenyDi-2 ethylpropane, where R' is theethyl' group crystallizes as awhitesolid which may be-filteredofi. It hasa melting point of 3. By this-method of hydrogenation, both the double bond and the ketone group in the startingmaterial are completely reduced (see Step II below). If desired, the product from. this hydrogenation may be converted into the corresponding phenolic product as Example I (see Step III below).

productionz 'oir 4.4"-dimethoxymsethyl chalcone (see. Step1 below)".

The chalcone as. produced: abovemay be re-- acted with. ethyl magnesium bromide in the; fol-.-

lowing manner: A solution. of. 8. grams of the chalcone -in 150. cc. of" dry ether; is added'islowly to a vigorously stirred: solution of 13.2.gramsof ethyl magnesium bromide in cc. of dry ether maintained at a temperature of --10-G. Stirring is eontinued for three hours at roomitemperaturef after which-the reaction mixture is. poured into. 500. cc. of ice. water containing '75. cc. of hydrochloric. acid. The ether layer is separated, the: aqueous solution extracted once with ether and; the ether solutions combined. After. evaporation. of" the ether. the residue. is; distilled under highvacuum. The distillate, 1,3-di-(prmethoxyphenyD-Z-ethyl pentanone-l-,. where R.- and R1 are. ethyl radicals, is: apale. yellow oil (see Step II; below. This; may: be further converted as follows:

10 grams of the ethylpentanone-as prepared'in. Step II is dissolved in 5.0 cc. of, absolute. alcohoL. 1 gram of a. copperechromium oxide, catalyst. added, and the mixture submittedgto hign pres.- sure hydrogenation at 220- and about. *atmose 'pheres pressureior 5 hoursinan appropriate ma? Pnonuc'rrorr or TYPE II Step-.11.

CHaO-OCHy-CH-CHOQCH; HOOCHwCHHOH -O-OH i y r y fi Any compound corresponding to the formula of Type II may be produced by followingthe basic outline of the process set 'forthabove, taking care in Step I to select the proper ketone for combination with the aldehyde.

EXAMPLE 3PRODUCTION OF. TYPE III Specific members of the series. illustrated by Type III may be prepared. in the followingv manner: The first step in this preparation is identical with the first step of Example 2, for the.

chine. After cooling, the hydrogen pressure is released and the solution removed from the hydrogenation apparatus and filtered. free of the catalyst. The alcohol is removed from the filtrate by evaporation and the product (see Step III below), which is 1,3-di(p-methoxypheny1) -2- ethylpentane, where Rand R1. are ethyl radicals.

is distilled at -175 C. under 1 mm. pressure. It is acoiorless liquid and may be converted into the corresponding phenolic compound as in Example I' (see Step IV'below) PRODUCTION or TYPE III Step I IV, until no more hydrogen is consumed. The 20 151 to a solution or ethyl magnesium bromide pregroups. The terminal groups may be in the pared from 5.0 grams of ethyl bromide and 1.2 ortho, meta or para position,'symmetrical or not. grams of magnesium turnings in anhydrous To produce terminal groups which are esters of ether. When the addition of the pentanone has inorganic acids, the phenolic terminal group is been completed, the reaction mixture is warmed condensed with an appropriate inorganic acid, on a steam bath for five hours after which it is such as phosphoric and, sulfonic acid. Ethyl poured into a mixture of 700 cc. ice water and magnesium bromide was used in each of the 100 cc. hydrochloric acid. The ether layer is sepforegoing reactions and excellent results are obarated, dried, and the ether removed. The resitained by its use, but it can be replaced by any due is subjected to vacuum distillation, and is alkyl magnesium halide except the fluoride.

obtained as a colorless oil. During this distilla- The compounds of thi in ti are prefertion, water is evolved, and the resulting unsatably administered by intramuscular injection and mated Product is -e y i P-methoXyshow the reaction of the ovarian follicular phenyD-heptene-2, where R1 and R2 are ethyl hormone.

groups. See StepIIIbelow. In the present specification and claims the The 4 ethyl-3,5-di(p-methoxyphenyl) hepsymbol tene-2 from the previous step is dissolved in '70 cc. glacial acetic acid and subjected to catalytic O I hydrogena ion as desc ibed in Example 4 Step sigmfies the benzene nucleus.

The dihydroxy compounds of this invention ixt t d th 1 m ure 1s hen decante from e cata yst, d1 have the general form luted with water, neutralized with sodium hydroxide, and the product extracted with ether. H0 R R R OH The ether is evaporated at room temperature, and

the remaining liquid distilled under reduced presl I I sure. The product thus obtained (see Step IV H E below) is a colorless oily liquid and is 4-ethylwhere R is selectedifrom the o p nsistin of 3,5-di(p-methoxyphenyl)-heptane, where R, R1 ydrogen and alkylf at least One Of Said and R2 are ethyl radicals, and may be converted R Substituents lbemg an l y calhe into the corresponding phenolic compound, as in hydroxy subs s may be in the ortho,

Example 1. or para position, and symmetrical or unsym- PRODUCTION or TYPE V Step I cmoOc a=o cmo-Oo-cmm CH;OOC-C=CH- 3OOH;

Step II cmoc-c=on ocm R,MgBr cmoOo-oH-cH-O-ocn. (l i. ll R1 in Step III cmoGc-cn-cn-O-oom RMgBr cH.0-C cH-ci1-oc11, g h R: B it; it:

StepIV cmoOc-cH-oH-O'ocm hydrogenation cmo-OcH-cH-oHO-ocn. t It. in t 1L. Ra Step V cum-Ocu-cn-cnOoom HOG-(FH-OH-CHOOH 1 2 1L4 I!!! R, R1 and R2 may be the same or different alkyl metrical. The invention also includes generically radicals. In general, any compound correspondthe ether and ester derivatives of these hydroxy ing to this type formula may be made by emcompounds. Either or both of the hydroxy subploying in Step I a ketone having the desired alkyl stituents may exist in the form of their functional radical, and in Steps II and III by employing derivatives-that is, ether and ester derivativesmagnesium compounds having the desired alkyl and those corresponding ether and ester comradicals. Mixtures of the racemates produced pounds may also be symmetrical or unsymmay be separated into the individual racemates, metrical.

e. g. by the method disclosed in the copending Further support for this generic statement, in application hereinabove referred to. addition to that already set forth, is provided by In the above reactions the terminal group was the following:

'methoxy, but it may be any alkoxy group, and Example 1 shows the production of may be obtained in the final product, by the use Ha Ha C2115 of appropriate starting compounds in Step I or I H by converting the OH groups in the final phe- HOO-d-Q-iQQH nolic compound. When a terminal acyl group is desired in the final compound it may be conveniently obtained by converting the OH groups By substituting meta-methoxyacetophenone in the final phenolic compound into RCOO for the para compound in Example 1 in column 3 and proceeding exactly as prescribed in that example, there is obtained (A) no EH11: 1%: (firm By substituting ortho-metli'oxyacetophenone for the para compound in Example 1 and proceeding otherwise exactly as in that example, there is obtained (B) HO Ha Hg C2H6 By substituting ortho-methoxybenzaldehyde for the para compound in Example 1 and n-propyl magnesium :bromide for ethyl magnesium bromide and proceeding otherwise exactly as in that example, there is obtained H2 H2 CaH1 HOOLM i Example 3 shows the production of H2 01H: C2115 HO i OH By substituting ortho-methoxybenzaldehyde for the para compound in Example 3 and otherwise proceeding exactly as in that example, there is obtained I i (D) 0H 03H, 01H: E0 o t t O it, t t

Example 4 shows the production of C2115 2 5 no t e t on G g, 51

By substituting ortho-methoxybenzaldehyde for the para compound and proceeding other-a wise exactly as shown in that example, there is obtained (E) on i a It I Example 5 shows the production of C Hs 02H; 02H;

Gea By substituting ortho-methoxylbenzaldehyde for the para compound and methyl magnesium iodide for ethyl magnesium bromide in Step 111 of that example and otherwise proceeding exactly as shown in Example 5, there is obtained I OH: ?z s (3215B G 0 l=![ i: ii

The above identified compounds (A) to (F) inclusive have been tested by subcutaneously injecting oil solutions thereof into castrated female rats and said compounds have been found to possess estrogenic activity comparable to. compounds shown in Examples 1 to 5 difiering from com- 12 pounds (A) to (F) inclusive'only in the position of the 0H groups.

By substituting in Example 4 the ortho-methoxy substituted compounds instead of the para substituted compounds, and otherwise proceeding exactly as in said example, there is obtained i i canooo-Qd-c-o-Goooom,

CaHi OgHa CaHu (5: 5 5

CzHs 02 5 Esters such as those listed above may-be prepared by any of the usual esterification methods. The following procedures are representative examples:

One gram of LS-di-(p-hydroxy phenyD-nhexane is dissolved in 5 cc. of benzoyl chloride and to this solution one drop of concentrated sulfuric acid is added. After standingseveral hours, 75 the mixture is diluted with water, made alkaline 13 with potassium hydroxide and extracted with ether. The ether solution is separated and after boiling off the ether, the residue is crystallized from alcohol. The dibenzoate is a white crystalline solid melting at 65 C.

One gram of 1,3-di- (p-hydroxy phenyl) -2-ethyl pentane is refluxed for four hours with a mixture of 10 cc. of acetic anhydride and 0.5 gram of fused sodium acetate. After dilution with water, the mixture is made alkaline with potassium hydroxide, and extracted with ether. The ether solution is separated, the ether boiled off and the residue distilled under high vacuum. It is a thick, pale yellow oil.

Three grams of palmityl chloride is added dropwise to a solution of 1 gram of 2,4-di-(p-hydroxy phenyl) -3-ethyl-hexane in 10 cc. of pyridine. After standing overnight, the reaction mixture is worked up in the usual manner. The dipalmitate, after crystallization from alcohol, is a white crystalline solid melting at 3638 C. All of the above esters have estrogenic properties.

As for the ether derivatives, the invention includes generically the monoor di-alkyl ether derivatives of the hydroxy compounds. Support Lf'OI' this generic statement is found in the f oregoing description, in which it is pointed out that instead of employing in Step I methoxy compounds, alkoxy compounds in general may be substituted, and that these alkoxy groups may be in the ortho, meta, or para position in relation to the other substituents, thus producing as a final step prior to hydrolysis, alkoxy or ether derivatives in general, symmetrical and unsymmetrical. Instead of producing alkoxy or ether derivatives by starting with appropriate alkoxy compounds, it is entirely possible to convert the final dihydroxy derivatives into their corresponding ethers by known etherification methods. Various illustrative specific examples of ether derivatives are shown; as follows:

i t? 031110 o-o-cOoolm OCaHo t 1 C4Hn0 C--(|3--C C2115 gH Hz E E3 As previously pointed out, there is an R substituent at one or more of the three carbon atoms of the propane bridge connecting the benzene radicals, and that R substituent is an alkyl radical in general, and those R substituents are the same or difierent alkyl radicals in general. Further support for these generic statements will be found in the following specific examples:

It is preferred that the number of carbon atoms in each of the alkyl substituents on the propane bridge be not greater than five, although the activity of such compounds having longer substituent chains has been proved by many experiments, numerous examples of which are found herein.

We claim:

1. Alpha, gamma-di(acyloxyphenyl) propanes in which each of the propane carbon atoms is substituted by a single alkyl group of less than six carbon atoms and in which the acyloxy groups are selected from organic acid radicals having from 1 to- 15 carbon atoms per radical.

2. 3-ethyl-2,4-di(p-acyloxyphenyl) hexane in which the acyloxy groups are selected fromorganic acid radicals having from 1 to '7 carbon atoms per radical.

3. 3-ethyl-2,4-di(p-acetoxyphenyl) hexane.

4. A racemic mixture of two stereoisomeric forms of the substance 3-ethyl-2,4-di(p-acyloxyphenyl) hexane in which the acyloxy groups are selected from organic acid radicals having from 1 to 7 carbon atoms per radical, said mixture having an estrogenic potency higher than the conglomerate of the eight possible stereoisomers of this substance.

RALPH C. TALLMAN. ALFRED H. STUART.

REFERENCES CITED The following references are of record in the file of this patent: 

